Interaction of eukaryotic translation initiation factor 4G with the nuclear cap-binding complex provides a link between nuclear and cytoplasmic functions of the m7 guanosine cap

In eukaryotes the majority of mRNAs have an m7G cap that is added cotranscriptionally and that plays an important role in many aspects of mRNA metabolism. The nuclear cap-binding complex (CBC; consisting of CBP20 and CBP80) mediates the stimulatory functions of the cap in pre-mRNA splicing, 3' end formation, and U snRNA export. As little is known about how nuclear CBC mediates the effects of the cap in higher eukaryotes, we have characterized proteins that interact with CBC in HeLa cell nuclear extracts as potential mediators of its function. Using cross-linking and coimmunoprecipitation, we show that eukaryotic translation initiation factor 4G (eIF4G), in addition to its function in the cytoplasm, is a nuclear CBC-interacting protein. We demonstrate that eIF4G interacts with CBC in vitro and that, in addition to its cytoplasmic localization, there is a significant nuclear pool of eIF4G in mammalian cells in vivo. Immunoprecipitation experiments suggest that, in contrast to the cytoplasmic pool, much of the nuclear eIF4G is not associated with eIF4E (translation cap binding protein of eIF4F) but is associated with CBC. While eIF4G stably associates with spliceosomes in vitro and shows close association with spliceosomal snRNPs and splicing factors in vivo, depletion studies show that it does not participate directly in the splicing reaction. Taken together the data indicate that nuclear eIF4G may be recruited to pre-mRNAs via its interaction with CBC and accompanies the mRNA to the cytoplasm, facilitating the switching of CBC for eIF4F. This may provide a mechanism to couple nuclear and cytoplasmic functions of the mRNA cap structure

A Description of an Improved Homodyne Laser Interferometer

First appearing on the commercial market in the mid 1980’s, diode-pumped, continuous-wave (cw) Nd:YAG lasers have more recently been used to obtain visible output, by the incorporation of frequency doubling optics in the laser cavity.The laser diode pumping of a Nd:YAG laser rod is selective and highly efficient, resulting in compact, high power, spatial mode lasers. Frequency-doubling processes are non-linear and lead to doubling TEM00 only of the high energy fundamental temporal mode, resulting in operation of the 532 nm laser in a single spatial and single longitudinal mode. The technology is rapidly advancing, and green lasers with energies of up to 1W could soon be available. The beam properties of the lasers described above are highly desirable in the field of interferometry, where such lasers are now in direct competition with the much larger Argon lasers, which have already been employed in high power interferometric systems. We describe here the performance of a modified Michelson interferometer [1–4], built to incorporate a 90 mW ADLAS 300 diode-pumped Nd:YAG laser. In previous versions of the Michelson interferometer, we have used HeNe lasers with a few milliwatts output, requiring mirror-quality surfaces on our samples. A 90 mW laser power enables us to make displacement measurements on metal surfaces with little or no preparation. The laser could also, of course, be used in other, more elaborate interferometer types, such as the confocal Fabry-Perot, which are better suited to industrial environments

The Met Office Unified Model Global Atmosphere 7.0/7.1 and JULES Global Land 7.0 configurations

We describe Global Atmosphere 7.0 and GlobalLand 7.0 (GA7.0/GL7.0), the latest science configurations of the Met Office Unified Model (UM) and the Joint UK Land Environment Simulator (JULES) land surface model developed for use across weather and climate timescales. GA7.0 and GL7.0 include incremental developments and targeted improvements that, between them, address four critical errors identified in previous configurations: excessive precipitation biases over India, warm and moist biases in the tropical tropopause layer (TTL), a source of energy non-conservation in the advection scheme and excessive surface radiation biases over the Southern Ocean. They also include two new parametrisations, namely the UK Chemistry and Aerosol(UKCA) GLOMAP-mode (Global Model of Aerosol Processes) aerosol scheme and the JULES multi-layer snow scheme, which improve the fidelity of the simulation and were required for inclusion in the Global Atmosphere/Global Land configurations ahead of the 6th Coupled Model Inter-comparison Project (CMIP6).In addition, we describe the GA7.1 branch configuration, which reduces an overly negative anthropogenic aerosol effective radiative forcing (ERF) in GA7.0 whilst maintaining the quality of simulations of the present-day climate. GA7.1/GL7.0 will form the physical atmosphere/land component in the HadGEM3–GC3.1 and UKESM1 climate model submissions to the CMIP6

Análisis de programas de mejora continua. Un estudio longitudinal en una empresa industrial

Las empresas han utilizado diversas herramientas que permiten que los operarios contribuyan al proceso de mejora continua. Entre las herramientas más usadas podemos destacar los sistemas de sugerencias tanto individuales como en grupo. En esta comunicación haremos un repaso de las principales características de ambos sistemas y los modos habituales de implantación. Nuestra ponencia pretende intentar responder a estas preguntas de investigación. ¿Qué resultados se derivan de la implantación de sistemas de sugerencias individuales o en grupo? ¿Cuál de los dos sistemas es más beneficioso para la empresa? ¿Qué problemas surgen durante el funcionamiento de estos programas? Para ello, analizaremos los datos de un caso de empresa industrial donde hemos recogido los datos históricos de 5 años de aplicación de un programa de mejora continua. Ambos programas han demostrado ser provechosos para la empresa, aunque las posibilidades de los sistemas de grupo parecen ser significativamente mayores

A gastrin transcript expressed in gastrointestinal cancer cells contains an internal ribosome entry site

As the hormone gastrin promotes gastrointestinal (GI) cancer progression by triggering survival pathways, regulation of gastrin expression at the translational level was explored. Sequence within the 5′ untranslated region of a gastrin transcript expressed in GI cancer cells was investigated, then cloned into a bicistronic vector upstream of firefly luciferase and transfected into a series of GI cancer cell lines. Firefly luciferase activity was measured relative to that of a cap-dependent Renilla luciferase. A gastrin transcript that was different from that described in Ensembl was expressed in GI cancer cells. Its transcription appears to be initiated within the region designated as the gene's first intron. In GI cancer cells transfected with the bicistronic construct, firefly luciferase activity increased 8–15-fold compared with the control vector, and there was a further induction of the signal (up to 25-fold) following exposure of the cells to genotoxic stress or hypoxia, suggesting that the sequence acts as an internal ribosome entry site. These data suggest that the gastrin transcript within GI cancer cells contains an internal ribosome entry site that may allow continued expression of gastrin peptides when normal translational mechanisms are inactive, such as in hypoxia, thereby promoting cancer cell survival

Effect of Manipulative Management Options on a Degraded Pasture within the Astrebla spp. Grazing Lands of Western Queensland

A reduction in Aslrebla spp., and an increase in less palatable and less desirable forbs (Malvastmm americanum Psoralea cinerea), was identified as a problem in some paddocks of the Astrebla grazing lands of central western Queensland\u27s clay soils. To determine possible rehabilitation for these pastures and assist the recovery of Astrebla spp,, a 2-year experiment was conducted near Muttaburra, Queensland (460 mm average annual rainfall), Six main treatments (nil, tine ripping, herbicide, slashing, fire, and a pioneering grass) were combined in a 6x2x2 split piot structure with grazed/not grazed and with additional Astrebla lappacea seed sown/not sown. Interim results showed that climate had a greater effect on the pasture than did the treatments. Stocking pressure was shown to be a powerful pasture manipulating option at the landholder\u27s disposal especially when considering ease of application, Tine cultivation was detrimental to Astrebla pastures. Atrazine significantly increased grass yield, with future implications for strategic grass seed production during moderate to high winter rainfalUlow summer rainfall years

Inhibition of Hypoxia-Inducible Factor-1α (HIF-1α) Protein Synthesis by DNA damage inducing agents

10.1371/journal.pone.0010522PLoS ONE55

Intravenous apoptotic spleen cell infusion induces a TGF-beta-dependent regulatory T-cell expansion.: Apoptosis and regulatory T cells

International audienceApoptotic leukocytes are endowed with immunomodulatory properties that can be used to enhance hematopoietic engraftment and prevent graft-versus-host disease (GvHD). This apoptotic cell-induced tolerogenic effect is mediated by host macrophages and not recipient dendritic cells or donor phagocytes present in the bone marrow graft as evidenced by selective cell depletion and trafficking experiments. Furthermore, apoptotic cell infusion is associated with TGF-beta-dependent donor CD4+CD25+ T-cell expansion. Such cells have a regulatory phenotype (CD62L(high) and intracellular CTLA-4+), express high levels of forkhead-box transcription factor p3 (Foxp3) mRNA and exert ex vivo suppressive activity through a cell-to-cell contact mechanism. In vivo CD25 depletion after apoptotic cell infusion prevents the apoptotic cell-induced beneficial effects on engraftment and GvHD occurrence. This highlights the role of regulatory T cells in the tolerogenic effect of apoptotic cell infusion. This novel association between apoptosis and regulatory T-cell expansion may also contribute to preventing deleterious autoimmune responses during normal turnover

The human insulin receptor mRNA contains a functional internal ribosome entry segment.

Regulation of mRNA translation is an important mechanism determining the level of expression of proteins in eukaryotic cells. Translation is most commonly initiated by cap-dependent scanning, but many eukaryotic mRNAs contain internal ribosome entry segments (IRESs), providing an alternative means of initiation capable of independent regulation. Here, we show by using dicistronic luciferase reporter vectors that the 5'-UTR of the mRNA encoding human insulin receptor (hIR) contains a functional IRES. RNAi-mediated knockdown showed that the protein PTB was required for maximum IRES activity. Electrophoretic mobility shift assays confirmed that PTB1, PTB2 and nPTB, but not unr or PTB4, bound to hIR mRNA, and deletion mapping implicated a CCU motif 448 nt upstream of the initiator AUG in PTB binding. The IR-IRES was functional in a number of cell lines, and most active in cells of neuronal origin, as assessed by luciferase reporter assays. The IRES was more active in confluent than sub-confluent cells, but activity did not change during differentiation of 3T3-L1 fibroblasts to adipocytes. IRES activity was stimulated by insulin in sub-confluent cells. The IRES may function to maintain expression of IR protein in tissues such as the brain where mRNA translation by cap-dependent scanning is less effective

The importance of the cellular stress response in the pathogenesis and treatment of type 2 diabetes

Organisms have evolved to survive rigorous environments and are not prepared to thrive in a world of caloric excess and sedentary behavior. A realization that physical exercise (or lack of it) plays a pivotal role in both the pathogenesis and therapy of type 2 diabetes mellitus (t2DM) has led to the provocative concept of therapeutic exercise mimetics. A decade ago, we attempted to simulate the beneficial effects of exercise by treating t2DM patients with 3 weeks of daily hyperthermia, induced by hot tub immersion. The short-term intervention had remarkable success, with a 1 % drop in HbA1, a trend toward weight loss, and improvement in diabetic neuropathic symptoms. An explanation for the beneficial effects of exercise and hyperthermia centers upon their ability to induce the cellular stress response (the heat shock response) and restore cellular homeostasis. Impaired stress response precedes major metabolic defects associated with t2DM and may be a near seminal event in the pathogenesis of the disease, tipping the balance from health into disease. Heat shock protein inducers share metabolic pathways associated with exercise with activation of AMPK, PGC1-a, and sirtuins. Diabetic therapies that induce the stress response, whether via heat, bioactive compounds, or genetic manipulation, improve or prevent all of the morbidities and comorbidities associated with the disease. The agents reduce insulin resistance, inflammatory cytokines, visceral adiposity, and body weight while increasing mitochondrial activity, normalizing membrane structure and lipid composition, and preserving organ function. Therapies restoring the stress response can re-tip the balance from disease into health and address the multifaceted defects associated with the disease